Mitochondrial Transfer from Bone Marrow Mesenchymal Stem Cells to Motor Neurons in Spinal Cord Injury Rats via Gap Junction
Mitochondrial Transfer from Bone Marrow Mesenchymal Stem Cells to Motor Neurons in Spinal Cord Injury Rats via Gap Junction
复制标题
线粒体通过间隙连接从骨髓间充质干细胞转移到脊髓损伤大鼠的运动神经元
DOI：
10.7150/thno.29400
复制
发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Wang, Lin-lin
中科院分区：
文献类型：
--
作者：
Li, Heyangzi;Wang, Chao;Wang, Lin-lin
近年来的研究表明，骨髓间充质干细胞（BMSCs）对脊髓损伤（SCI）后神经元的凋亡具有保护作用，但其保护作用的机制尚不清楚。本研究发现骨髓间充质干细胞的线粒体可以移植到受损的运动神经元中，并检测移植后的功能改善情况。方法：将原代大鼠BMSCs与氧糖剥夺（OGD）损伤的VSC4.1运动神经元或原代皮层神经元共培养。流式细胞仪检测骨髓间充质干细胞线粒体向神经元的转移。细胞外液流分析检测神经元的生物能量学特征。还测量了细胞活力和凋亡。骨髓基质细胞和分离的线粒体移植到脊髓损伤大鼠。采用TdT介导的dUTP缺口末端标记法检测腹角神经元凋亡。免疫组化和Western blotting检测蛋白表达。透射电镜观察髓鞘再生情况。BBB评分用于评估运动功能。结果：MitoTracker-Red标记的骨髓间充质干细胞线粒体可转移至OGD损伤的神经元。差距连接细胞间通讯（gap junction intercellular communication，GJIC）增强剂维甲酸可增加BMSCs向神经元的线粒体转运量，而GJIC抑制剂18β-大黄酸可减少线粒体转运量。线粒体的内化改善了OGD后运动神经元的生物能量学特征，减少了细胞凋亡并促进了细胞存活。此外，线粒体和骨髓间充质干细胞移植到损伤的脊髓促进SCI大鼠运动功能的恢复。结论：据我们所知，这是第一个证明BMSCs通过GJIC将线粒体转移到受损的神经元来保护脊髓损伤的证据。本研究为脊髓损伤患者的线粒体移植治疗提供了新的思路。
Recent studies have demonstrated that bone marrow mesenchymal stem cells (BMSCs) protect the injured neurons of spinal cord injury (SCI) from apoptosis while the underlying mechanism of the protective effect of BMSCs remains unclear. In this study, we found the transfer of mitochondria from BMSCs to injured motor neurons and detected the functional improvement after transplanting. Methods: Primary rat BMSCs were co-cultured with oxygen-glucose deprivation (OGD) injured VSC4.1 motor neurons or primary cortical neurons. FACS analysis was used to detect the transfer of mitochondria from BMSCs to neurons. The bioenergetics profiling of neurons was detected by Extracellular Flux Analysis. Cell viability and apoptosis were also measured. BMSCs and isolated mitochondria were transplanted into SCI rats. TdT-mediated dUTP nick end labelling staining was used to detect apoptotic neurons in the ventral horn. Immunohistochemistry and Western blotting were used to measure protein expression. Re-myelination was examined by transmission electron microscope. BBB scores were used to assess locomotor function. Results: MitoTracker-Red labelled mitochondria of BMSCs could be transferred to the OGD injured neurons. The gap junction intercellular communication (GJIC) potentiator retinoid acid increased the quantity of mitochondria transfer from BMSCs to neurons, while GJIC inhibitor 18β glycyrrhetinic acid decreased mitochondria transfer. Internalization of mitochondria improved the bioenergetics profile, decreased apoptosis and promoted cell survival in post-OGD motor neurons. Furthermore, both transplantation of mitochondria and BMSCs to the injured spinal cord improved locomotor functional recovery in SCI rats. Conclusions: To our knowledge, this is the first evidence that BMSCs protect against SCI through GJIC to transfer mitochondrial to the injured neurons. Our findings suggested a new therapy strategy of mitochondria transfer for the patients with SCI.